Safety in cell and tissue culture Safety in cell and tissue eult ure Edited by G. Stacey National Institute for Biological Standards and Control South Mimms, Hertfordshire, UK A. Doyle European Collection of Cell Cultures Centre for Applied Microbiology and Research Salisbury, Wiltshire, UK and P. Hambleton Centre for Applied Microbiology and Research Salisbury, Wiltshire, UK SPRINGER SCIENCE+BUSINESS MEDIA, B.V. A c.I.P. Catalogue record for this book is available from the Library of Congress ISBN 978-94-010-6061-5 ISBN 978-94-011-4916-7 (eBook) DOI 10.1007/978-94-011-4916-7 Printed an acid-free paper AH Rights Reserved © 1998 Springer Science+Business Media Dordrecht Originally published by Kluwer Academic Publishers in 1998 No part of material protected by this copyright notice may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording or by any information storage and retrieval system, without written permis sion from the copyright owner. Contents Colour plates appear between pp. 118 and 119 List of Contributors vii Preface ix 1 Source materials 1 Glyn Stacey, Alan Doyle dnd David Tyrrell 2 Cell biology aspects of safety in cell culture 26 Trevor Littlewood 3 Viral contamination of cell cultures 49 Alan Jennings 4 Laboratory practice 64 Bruce Jones 5 Planning and design of a cell and tissue culture laboratory 87 Christopher Morris 6 Quality control and validation 102 Alan Doyle and Glyn Stacey 7 Containment facilities: design, construction and working practices 116 John Benbough and B. Andrew Curran 8 Scale-up of animal cell culture systems 135 Bryan Griffiths and Wolfgang Noe 9 Production and containment of bioreactor processes 155 Geoffrey Leaver 10 Risk assessment 173 Heather Sheeley 11 Safety aspects of genetic modification procedures 189 Caroline MacDonald 12 International guidelines for safe packaging and transport of biological materials 205 Christine Rohde and Dieter Claus vi Contents Appendixes to Chapter 12: A Addresses of relevant international organizations 223 B Articles 118-120 of the Detailed Regulations of the 223 UPU Convention C Countries with import and export restrictions for NPBS and 224 IPBS for national postal services D Transportation and Packaging (Numerical Order) 226 E Suppliers of Certified Transport Containers 232 Appendix A: Classification of Microorganisms 233 Appendix B: Containment Level for Cell Culture 235 Index 236 Contributors John E. Benbough Biological Investigations Unit, Centre for Applied Microbiology and Research, Salisbury, Wiltshire, UK Dieter Claus DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg lB, Braunschweig, Germany B. Andrew Curran Centre for Applied Microbiology and Research, Salisbury, Wiltshire, UK Alan Doyle European Collection of Cell Cultures, Centre for Applied Microbiology and Research, Salisbury, Wiltshire, UK Bryan J. Griffiths Bourne Gardens, Porton, Wiltshire, UK Peter Hambleton Centre for Applied Microbiology and Research, Salisbury, Wiltshire, UK Alan Jennings Centre for Applied Microbiology and Research, Salisbury, Wiltshire, UK Bruce P.e. Jones National Institute for Biological Standards and Control, South Mimms, Hertfordshire, UK Geoffrey A. Leaver AEA Technology, Bio-Sciences, Harwell, Oxon, UK Trevor D. Littlewood Imperial Cancer Research Fund London, Lincoln's Inn Fields, London, UK Caroline MacDonald Department of Biological Sciences, University of Paisley, UK viii Contributors Christopher B. Morris The Wellcome Trust Centre for Human Genetics, Oxford, UK Wolfgang Noe Department of Biotechnical Production, Dr Karl Thomae GmbH, Biberach an der Riss, Germany Christine Rohde DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg lB, Braunschweig, Germany Heather Sheeley Centre for Applied Microbiology and Research, Salisbury, Wiltshire, UK Glyn Stacey National Institute for Biological Standards and Control, South Mimms, Hertfordshire, UK David Tyrrell Centre for Applied Microbiology and Research, Salisbury, Wiltshire, UK Preface It is now more than half a century since animal cells first came into regular use in the laboratory. Instances of laboratory acquired infection and con tamination of therapeutic products, derived from the use of animal cell cultures are rare. The use of animal cells, in addition to an established role in the production of vaccines and therapeutic proteins, has many new medical applications including gene therapy, tissue engineering and cell therapy. Furthermore, C;ldvances in molecular and cell biology are enabling rapid development and application of these technologies and the development of new and more sensitive methods, such as nucleic acid amplification, for the characterisation of cells and the detection of adven titious agents. However, it is clear that there is no room for complacency in this field and the recent expansion in the use of animal cells in the manufacture of medical products and the development of new biological assays for diagnostic and pharmaco-toxicological screening, underlines the need for vigilance regarding the correct and safe use of animal cells as substrates. This book is therefore very timely and should prove to be a highly valuable text, finding a wider audience beyond those with respon sibility for laboratory safety. The book guides the reader from fundamental cell biology issues and the establishment of new in vitro methods, through testing and validation of cell lines and on to issues in the use of animal cells in manufacturing processes. There is constant reference to national and international guide lines which provides a solid framework. However, no legislation can replace the practical experience which is delivered in this book in order to ease the path of new cell substrates and in vitro techniques through research to clinical exploitation by avoiding over-regulation and un necessary bureaucracy. This is a unique publication providing some of the best available biosafety guidance for newcomers to the practice of cell and tissue culture. It offers an extremely valuable reference source which, if used in combina tion with appropriate training and monitoring procedures, will enhance the safety of laboratory work and biological products alike. Dr. Geoffrey C. Schild National Institute for Biological Standards and Control South Mimms, UK CHAPTER 1 Source materials Glyn Staceyl, Alan Doyli and David Tyrrelz2 1.1 INTRODUCTION When assessing the requirements and implications of a project involving cell and tissue culture, it is important to determine at the outset what type of culture system will fit the purpose, and whether the work is 'closed', with material not made available to third parties. These fundamental factors will determine the necessary control, containment and testing procedures which must be applied to the work. Other chapters will deal with these issues in detail, but in this chapter strategic approaches to selecting safe cell culture systems, and more specifically the hazards of raw materials, will be addressed. A key issue in handling animal cell cultures is that they may carry and possibly support the growth of microorganisms, primarily viruses, which are dependent on the biochemical machinery of animal cells to facilitate their replication. Where virus replication occurs and persists in vitro there may be a significant infectious hazard for laboratory workers, ancillary staff and recipients of material derived from the original cells, including patients treated with therapeutic agents derived from animal cells. At this point it is worthwhile to review the different types of in vitro cell culture available to the researcher and what their comparative values may be. Virtually all cell and tissue culture approaches can be included within certain basic definitions [1,2] as follows: • organ cultures: isolated organs and functional tissue, e.g. tissue slices; • primary cells: cells derived directly from animal tissue and cultured without passage; • finite cell lines: cells derived from normal tissue which are capable of replication and passage in vitro; such cultures may become adequately characterized to achieve the status of a 'cell strain', e.g. MRC-S, Wl-38; • continuous cell lines: cell cultures which appear to have the property of indefinite passage, e.g. CHO, HeLa. INational Institute for Biological Standards and Control, South Mimms, UK; 2Centre for Applied Microbiology and Research (CAMR), Salisbury, UK 2 G. Stacey, A. Doyle and D. Tyrrell Finite cell lines have been used to great effect over many years but gener ally represent a limited range of cell types (Le. mainly fibroblasts) and have a limited lifespan. The use of continuous cell lines overcomes these two limitations. However, these are often derived from abnormal tu mours or mutants (produced by the effects of toxic chemicals or radiation) which may not express the characteristics typical of the original tissue. In addition, even cell lines expressing such characteristics may lose them on repeated passage, although this phenomenon may be avoided by appro priate cell banking procedures (see below). Growth of animal cells on treated culture surfaces (e.g. collagen) or in the presence of certain bioactive substances (e.g. retinoic acid) may enhance the differentiated characteristics of cells in culture. However, such conditions are in general mutually exclusive with proliferation and are therefore not appropriate when expansion of the culture is desired. There are cases of certain recombinant cells which carry genetic constructs, involved in the stimula tion of cell proliferation, for which expression can be regulated. This effect can be mediated by temperature shift (e.g. ts genes) or treatment of the cells with modified biological inducers or inhibitors of the recombinant proliferation mechanism (e.g. tamoxifen) [3]. Spontaneous transformation in vitro has often been described but, as its name suggests, is little understood as a process. This phenomenon is therefore a cause for concern in risk assessment, but in examples such as the Vervet monkey kidney cell line Vero, a commonly used vaccine substrate, this has not prevented regulatory approval. However, in cases of spontaneous transformation it is important to exclude the possibility of cross-contamination of the original culture by a continuous cell line. The discovery of widespread cross-contamination of cell lines by the HeLa cells in the 1970s demonstrated the potential for such incidents to go unrecognized in tissue culture laboratories [4]. The proliferative capacity and relative stability of finite and continuous cell lines enable the preparation of bulk stocks, called 'cell banks', which can be quality controlled. Thus the availability of reliable and repro ducible samples for widespread distribution and long-term preservation enhances the level of standardization which can be achieved in research and industry. The need for such standardized cell cultures was recog nized in the 1960s when it was discovered that the Salk poliovaccine was contaminated with SV 40 virus which came from the monkey kidney cells in which the vaccine was produced [5]. The acceptability and regulatory philosophy relating to the use of animal cells in the production of biologicals has been reviewed by Petricciani [6] and more recently ad dressed by the World Health Organisation (WHO) Expert Committee on Biological Standardisation [7]. Detailed approaches to risk assessment relating to these different culture types are described in Chapter 10, but here we will deal with more general issues. In choosing an in vitro model system, a number of